Immunohistology for Quantification of Normal Bone Marrow Lymphocyte Subsets

Thaler, Josef; Greil, Richard; Dietze, Otto; Huber, H.; Kaiserling, E.; Horny, Hans‐Peter; Engst, U.; Walz, Rainer

doi:10.1111/j.1365-2141.1989.tb00307.x

Cited by 24 publications

(13 citation statements)

References 6 publications

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“…In the present study, however, the T and the B subsets were present in equivalent percentages in adults, in agreement with the results obtained in B-5 fixed decalcified BM biopsies reported by O'Donnel et al (32). However, the percentages of lymphoid subsets reported by immunohistochemical studies (21,32,39) are relative to total nucleated cells in BM or to the analyzed area (32), and comparison with those results obtained by flow cytometry is inaccurate. Considering only the previous studies of BM by flow cytometry, our results showed higher percentages of both T and B cells, even compared with data obtained from biopsies (9).…”

Section: Discussionsupporting

confidence: 61%

Age-related changes of lymphocyte subsets in normal bone marrow biopsies

et al. 1998

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Section: Discussionsupporting

confidence: 61%

Age-related changes of lymphocyte subsets in normal bone marrow biopsies

et al. 1998

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“…A putative positive staining of a few histiocytic cells was recognized during morphometric evaluation of the bone marrow samples, but not regarded in our calculations. In confirmation and extension to previous findings (14,33), morphometric quantification on paraffin sections derived from our control group of patients showed a similar ratio (4:1) between CD45RO ϩ and CD20 ϩ lymphocytes in hematopoietic tissue (Table 2). On the other hand, a more comprehensively conducted comparison between normal and CML bone marrow specimens has to regard cellularity (i.e., the amount of hematopoiesis).…”

Section: Discussionsupporting

confidence: 91%

“…First of all, a conflict of opinion still persists regarding the exact number or ratio of T-and B-lymphocytes in the normal bone marrow. According to immunohistologic analysis of bone marrow frozen sections, the ratio is about 9% T-lymphocytes (2.7% CD4 ϩ and 6.3% CD8 ϩ ) to 1.5% (CD22 ϩ ) B-lymphocytes (33). This T-to B-cell ratio is generally comparable with the CD45R0-and CD20-reactivity of lymphocyte subsets in paraffin-embedded bone marrow trephine biopsies amounting between 4:1 and 5:1 (14).…”

Section: Discussionmentioning

confidence: 70%

“…Although immunohistochemical investigations of lymphocyte subsets can be easily performed on cryostat sections, this involves technical problems that generally have prevented the introduction of such methods into routine practice of hematopathology (32,33). This shortcoming has also significantly impaired a more elaborate evaluation of a large number of bone marrow specimens.…”

Section: Discussionmentioning

confidence: 99%

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Therapy-related Changes of CD20+ and CD45RO+ Lymphocyte Subsets in Chronic Myeloid Leukemia (CML): An Immunohistochemical and Morphometric Study on Sequential Trephine Biopsies of the Bone Marrow

et al. 2000

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Little information exists about the amount of CD45RO ؉ -T-and CD20 ؉ -B-lymphocytes in the bone marrow of patients with Philadelphia chromosome-positive chronic myelogenous leukemia (Ph 1؉ -CML) at presentation or regarding corresponding changes during therapy. On the other hand, quantification of this cell compartment seems to be imperative for two reasons: first, the presumed association of immunocompetent lymphocyte subsets in the expansion of the leukemic cell clone; and second, a speculated relationship with the complex generation of myelofibrosis. Therefore, an immunohistological and morphometric study was performed on 219 representative trephine biopsies of the bone marrow derived from 70 patients with repeated examinations during the course of Ph 1؉ -CML. For the identification of the different lymphocyte populations, the monoclonal antibodies UCHL-1 (CD45R0) and L26 (CD20) were applied on formaldehyde-fixed and decalcified specimens. In comparison to a control group and calculated per hematopoietic cells, the CML bone marrow showed about a 50% decrease in the total amount of lymphocytes. Determination of CD45RO ؉ and CD20 ؉ subsets revealed a significant enhancement during treatment. Because of the different intervals (range, 10 to 25 mo) between first and last biopsy in the various therapeutic groups, results had to be modified by considering dynamic features. This calculation included changes of the lymphocyte subpopulations related to time. Contrasting the CD45RO ؉ lymphocytes, a relevant increase in the CD20 ؉ subset could be observed after interferon-␣ treatment or corresponding combination regimens. No significant correlations were found between fiber density at onset (first biopsy) or development of fibrosis and lymphocyte proliferations in the course of CML. Our results are in keeping with the finding that a proper immune response consistent with an increased lymphocyte growth seems to be associated with a regression of the clonally-transformed cell population. Opposed to a repeatedly discussed pathomechanism, we failed to demonstrate any quantitative relationships between the extent of lymphocyte proliferations and occurrence or progression of myelofibrosis.

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“…These studies relied on BM donations from sibling donors, including a few children, or on trephine biopsies obtained during surgery or from volunteers. A major finding was that BM contains increased numbers of CD8+ T cells compared to peripheral blood, resulting in lower CD4/CD8 cell ratios [25,27,28,29]. Among BM lymphocytes upon diagnosis of ALL, we found highly variable CD4/CD8 ratios with a median of 1.22, slightly above that reported for normal BM.…”

Section: Discussionmentioning

confidence: 73%

High Proportions of CD4+ T Cells among Residual Bone Marrow T Cells in Childhood Acute Lymphoblastic Leukemia Are Associated with Favorable Early Responses

Lustfeld

Altvater²,

Ahlmann³

et al. 2013

Acta Haematol

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Residual nonmalignant T cells in the bone marrow of patients with acute leukemias may be involved in active immune responses to leukemic cells. Here, we investigated the phenotypic signature of T cells present at diagnosis in 39 pediatric patients with acute lymphoblastic leukemia (ALL) treated within standardized ALL-BFM study protocols. Previously described age associations of lymphocyte subpopulations in the peripheral blood of healthy children were reproduced in leukemic bone marrow. Analysis of individual lymphocyte parameters and risk-associated variables using univariate linear regression models revealed a correlation of higher CD4/CD8 ratios at diagnosis with a favorable bone marrow response on day 15. Separate analysis of CD4+ cells with the CD4+CD25^hiFoxP3+ T_reg cell phenotype showed that the association was caused by non-T_reg CD4+ cells. The association of higher CD4/CD8 ratios with a favorable bone marrow response on day 15 of treatment persisted in a cohort extended to 69 patients. We conclude that CD4+ non-T_reg cells in leukemic bone marrow at diagnosis may have a role in early response to treatment. Prospective analysis of the CD4/CD8 ratio in a large cohort of pediatric patients is now needed. Moreover, future experiments will establish the functional role of the individual T cell subsets in immune control in pediatric ALL.

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Immunohistology for Quantification of Normal Bone Marrow Lymphocyte Subsets

Cited by 24 publications

References 6 publications

Age-related changes of lymphocyte subsets in normal bone marrow biopsies

Age-related changes of lymphocyte subsets in normal bone marrow biopsies

Therapy-related Changes of CD20+ and CD45RO+ Lymphocyte Subsets in Chronic Myeloid Leukemia (CML): An Immunohistochemical and Morphometric Study on Sequential Trephine Biopsies of the Bone Marrow

High Proportions of CD4+ T Cells among Residual Bone Marrow T Cells in Childhood Acute Lymphoblastic Leukemia Are Associated with Favorable Early Responses

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